Comparison of two gonadotropin-releasing hormone agonist suppression protocols for in vitro fertilization in young patients with low body mass index.

OBJECTIVE: To evaluate two different gonadotropin-releasing (pituitary downregulating) hormones used in in vitro fertilization (IVF) on the live birth rate in young patients with low body mass index (BMI) undergoing their first IVF cycle. METHODS: In a retrospective study in a single public medical center, 555 long gonadotropin-releasing hormone agonist (GnRH-a) protocols were compared with 431 prolonged GnRH-a protocols between 2016 and 2018. All analyses were performed using the SPSS version 22.0. The primary measured outcome was live birth rate. RESULTS: Compared with the long protocol, the prolonged protocol required more doses of gonadotropin and a longer duration of ovarian stimulation. Lower levels of serum luteinizing hormone and serum estrogen were detected on the day of chorionic gonadotropin administration, and a lower fertilization rate was found in the prolonged protocol. Although more oocytes were retrieved and more frozen embryos were recorded in the prolonged protocol, the live birth rate per fresh cycle was comparable between the two protocols (P = 0.057). The incidence of ovarian hyperstimulation syndrome was higher in the prolonged protocol group. In the subgroup of women with antral follicle count (AFC) of 12 or less, there was no difference in the live birth rate between the two protocols (P = 0.688). However, for women with AFC > greater than 12, the prolonged protocol was still a positive predictor of live birth rate. The odds ratio was 1.73 (95% confidence interval 1.04-2.89). CONCLUSION: The prolonged protocol might not increase the live birth rate in women with low BMI who are undergoing their first IVF cycle. However, for women with AFC greater than 12, a prolonged protocol could be a good choice to improve the live birth rate.

Serum Metabolomic Signature Predicts Ovarian Response to Controlled Stimulation.

In in vitro fertilization (IVF), it is meaningful to find novel biomarkers predicting ovarian response in advance. The aim of the study was to identify serum metabolomics predicting ovarian response after controlled ovarian stimulation (COS). Blood samples collected at the start of pituitary downregulation and on the fifth day after COS using Liquid chromatography-tandem mass spectrometry (LC-MS/MS) methods were analyzed to quantify metabolites. Demographic data were calculated with SPSS version 22.0 software. Multivariate statistics were used to analyze metabolomics dataset. A receiver operating characteristic (ROC) curve was used to evaluate the diagnostic model. Analyses revealed 50 different metabolomics between the pre- and post-COS groups. Compared with baseline, amino acids increased significantly following COS. At baseline, acetylglycine was more abundant in FOI<1 group, while glycine and lipids increased in FOI≥1 group. After COS, glycine, N-acetyl-L-alanine, D-alanine, and 2-aminomuconic acid were higher in those with FOI≥1, but L-glutamine was abundant in FOI<1. ROC curves indicated that combination of glycine, acetylglycine, and lipids predicts different responses to COS (AUC=0.866). Serum metabolism might reflect the response to ovarian stimulation. Higher glycine and PC may be a good predictor for response to COS.

The interval between insemination and ovulation predicts outcome after intrauterine insemination with donor sperm (IUI-D).

OBJECTIVE: To identify whether the time interval from insemination to ovulation (I-O interval) affects outcome after intrauterine insemination with donor sperm (IUI-D). METHODS: A retrospective study was conducted in a public assisted reproductive medicine center between January, 2014 and December, 2016 in Xi'an, China. The data were collected from the medical records and generalized estimating equations (GEEs) were used to evaluate the effects of various variables on IUI outcome. RESULTS: A total of 2091 IUI-D cycles from 1165 couples were included in this study. Multiple predictors were identified for (live birth rate) LBR. The I-O interval was the predictor for LBR. An I-O interval ≥19 h significantly decreased CPR (odds ratio [OR], 95% confidence interval [CI]: 0.29, 0.17-0.48) and LBR (OR, 95% CI: 0.32, 0.19-0.55). The presence of at least two follicles ≥18 mm on ovulation day significantly increased the LBR (OR, 95%CI: 1.27, 1.01-1.60). Women aged 35 years and older had a significantly decreased LBR (OR, 95% CI: 0.61, 0.38-0.98). CONCLUSION: The I-O interval, a new prognostic factor, in combination with the woman's age and number of mature follicles, can predict the outcome after IUI-D. IUI-D is best performed within 19 h of I-O interval for a higher probability of clinical pregnancy and live birth.

Evaluation of Ovarian Reserve Tests and Age in the Prediction of Poor Ovarian Response to Controlled Ovarian Stimulation-A Real-World Data Analysis of 89,002 Patients.

Aims: This study aimed to explore the value of ovarian reserve tests (ORTs) for predicting poor ovary response (POR) and whether an age cutoff could improve this forecasting, so as to facilitate clinical decision-making for women undergoing in vitro fertilization (IVF). Methods: A retrospective cohort study was conducted on poor ovary response (POR) patients using real-world data from five reproductive centers of university-affiliated hospitals or large academic hospitals in China. A total of 89,002 women with infertility undergoing their first traditional ovarian stimulation cycle for in vitro fertilization from January 2013 to December 2019 were included. The receiver operating characteristic (ROC) curve was performed to estimate the prediction value of POR by the following ORTs: anti-Mullerian hormone (AMH), antral follicle count (AFC), basal FSH (bFSH), as well as patient age. Results: In this retrospective cohort, the frequency of POR in the first IVF cycle was 14.8%. Age, AFC, AMH, and bFSH were used as predicting factors for POR, of which AMH and AFC were the best indicators when using a single factor for prediction (AUC 0.862 and 0.842, respectively). The predictive values of the multivariate model included age and AMH (AUC 0.865), age and AFC (AUC 0.850), age and all three ORTs (AUC 0.873). Compared with using a single factor alone, the combinations of ORTs and female age can increase the predictive value of POR. Adding age to single AMH model improved the prediction accuracy compared with AMH alone (AUC 0.865 vs. 0.862), but the improvement was not significant. The AFC with age model significantly improved the prediction accuracy of the single AFC model (AUC 0.846 vs. 0.837). To reach 90% specificity for POR prediction, the cutoff point for age was 38 years old with a sensitivity of 40.7%, 5 for AFC with a sensitivity of 55.9%, and 1.18 ng/ml for AMH with a sensitivity of 63.3%. Conclusion: AFC and AMH demonstrated a high accuracy when using ROC regression to predict POR. When testing is reliable, AMH can be used alone to forecast POR. When AFC is used as a prediction parameter, age is suggested to be considered as well. Based on the results of the cutoff threshold analysis, AFC ≤ 5 and AMH ≤ 1.18 ng/ml should be recommended to predict POR more accurately in IVF/ICSI patients.

Association of IL-6 -174G>C (rs1800795) polymorphism with cervical cancer susceptibility.

Interleukin-6 (IL-6) is a multifunctional cytokine that has been implicated in the etiology of cancer. Several case-control studies have been conducted to assess the association of IL-6 -174G>C (rs1800795) polymorphism with the risk of cervical cancer, yet with conflicting conclusions. To derive a more precise estimation of the relationship, we performed this meta-analysis updated to June 2018. A total of seven original publications were identified covering IL-6 -174G>C (rs1800795) polymorphism. Odds ratios (ORs) and 95% confidence intervals (CIs) were used to assess the relationship strengths. Statistically significant relationship was observed between IL-6 -174G>C polymorphism and cervical cancer risk (OR = 0.61, 95% CI: 0.40-0.94 for GG vs. CC, and OR = 0.77, 95% CI: 0.64-0.93 for G vs. C). Moreover, the significant association was found among Asians (OR = 0.46, 95% CI: 0.29-0.75 for GG vs. CC, and OR = 0.70, 95% CI: 0.57-0.89 for G vs. C); hospital-based subgroup (OR = 0.53, 95% CI: 0.38-0.72 for GG vs. CC, and OR = 0.73, 95% CI: 0.61-0.87 for G vs. C); and Hardy-Weinberg equilibrium ≤0.05 (OR = 0.56, 95% CI: 0.37-0.86 for GG vs. GC, and OR = 0.66, 95% CI: 0.47-0.93 for G vs. C). This meta-analysis showed the evidence that the IL-6 -174G>C polymorphism was a low-penetrance susceptibility variant for cervical cancer. Further large-scale case-control studies are needed to confirm these results.

[The rate of morphologically normal sperm does not affect the clinical outcomes of conventional IVF in patients with one retrieved oocyte].

OBJECTIVE: To investigate the influence of the rate of morphologically normal sperm (MNS) on the clinical outcomes of conventional in vitro fertilization (IVF) in patients with one retrieved oocyte. METHODS: From January 2013 to January 2015, a total of 256 couples with one retrieved oocyte underwent conventional IVF in our center. According to the rate of MNS, the patients were divided into two groups: MNS < 4% (134 cycles) and MNS ≥ 4% (122 cycles). We compared the rates of no transferrable embryo cycles, fertilization, cleavage, normal fertilization, abnormal fertilization, high-quality embryo and transferrable embryo between the two groups. A total of 75 fresh embryo transfer cycles were performed, 43 in the MNS < 4% group and the other 32 in the MNS ≥ 4% group. We also compared the rates of implantation, clinical pregnancy and abortion between the two groups. RESULTS: There were no statistically significant differences between the two groups in the rates of no transferrable embryo cycles, fertilization, cleavage, normal fertilization, abnormal fertilization, high-quality embryo and transferrable embryo (P > 0.05). The rates of implantation, clinical pregnancy and abortion exhibited no remarkable differences either in the fresh embryo transfer cycles between the two groups (P > 0.05). CONCLUSION: The rate of MNS does not affect the clinical outcomes of conventional IVF in patients with one retrieved oocyte.

[Association of fertilization strategy and embryo transfer time with the incidence of ectopic pregnancy].

OBJECTIVE: To investigate the correlation of the fertilization strategy and embryo transfer (ET) time with the incidence of ectopic pregnancy. METHODS: We selected 3,331 fresh and 2,706 frozen-thawed ET cycles for the patients undergoing in vitro fertilization (IVF) and intracytoplasmic sperm injection (ICSI). The fresh transfers included 2 546 IVF-ET and 785 ICSI-ET cycles and 2,220 day-3 embryo and 1,111 day-5 blastocyst transfers, while the frozen-thawed transfers included 2,080 IVF-ET and 626 ICSI-ET cycles and 741 day-3 embryo and 1 965 day-5 or -6 blastocyst transfers. We compared the incidence rate of ectopic pregnancy associated with different fertilization strategies and ET time. RESULTS: The incidence rate of ectopic pregnancy was 1. 41% (36/2 546) in the IVF-ET cycles and 3.44% (27/785) in the ICSI-ET cycles of the fresh transfers, significantly lower in the IVF-ET than in the ICSI-ET cycles (P < 0.01), and it was 1.01% (21/2,080) in the IVF-ET cycles and 0.80% (5/626) in the ICSI-ET cycles of the frozen-thawed transfers, with no remarkable difference between the two groups (P > 0.05). The IVF-ET and ICSI-ET cycles included 2,220 fresh day-3 (F-D3) embryos, 1,111 F-D5 blastocysts, 741 frozen-thawed day-3 (T-D3) embryos, and 1,965 T-D5/6 blastocysts. The incidence rate of ectopic pregnancy was 1.71% (n = 38) in the F-D3, 2.25% (n = 25) in the F-D5, 1.35% (n = 10) in the T-D3, and 0.81% (n = 16) in the T-D5/6 group, respectively, significantly lower in the T-D5/6 than in the other three groups (P < 0.05). CONCLUSION: The incidence rate of ectopic pregnancy is associated with fertilization strategies, which is significantly lower in frozen-thawed than in fresh embryo transfers.

[Intracytoplasmic sperm injection does not improve the clinical outcomes of the males with 100% teratozoospermia].

OBJECTIVE: To investigate whether intracytoplasmic sperm injection (ICSI) can improve the clinical outcomes of the male patients with 100% teratozoospermia. METHODS: We retrospectively analyzed the clinical data of 152 couples undergoing in vitro fertilization-embryo transfer (IVF-ET), including 75 cycles of IVF and 77 cycles of ICSI. We compared the rates of normal fertilization, high-quality embryos, transferrable embryos, implantation, clinical pregnancy, and abortion between the two groups. RESULTS: In the 100% teratozoospermia patients the number of transferrable embryos was significantly lower in the IVF than in the ICSI group (78.91% vs 84.92%, P < 0.05), while the rates of normal fertilization and implantation were higher in the former than in the latter (60.26% vs 57.87% and 48.00% vs 39.55%, both P > 0.05). There were no statistically significant differences between the two groups in the female age, Gn days, Gn dose, BMI, infertility duration, endometrial thickness, and basal serum FSH and E2. CONCLUSION: ICSI cannot improve the clinical outcomes of the male patients with 100% teratozoospermia.

[Influence factors on the cryosurvival rate of post-thaw spermatozoa from sperm donors].

OBJECTIVE: To investigate the influence of seasons, blood types and semen parameters on the cryosurvival rate of frozen-thawed spermatozoa from sperm donors. METHODS: We retrospectively analyzed the influence of seasons, blood types, abstinence period, semen volume, sperm morphology, and pre-freeze sperm motility and concentration on the cryosurvival rate of frozen-thawed sperm in 4 088 semen samples from Shaanxi Human Sperm Bank. RESULTS: The cryosurvival rate of the post-thaw sperm was correlated negatively with the progressive motility of the pre-freeze sperm (r = -0.262, P < 0.01), but positively with pre-freeze sperm concentration (r = 0.247, P < 0.01), and it was significantly higher in the 6-day abstinence group ([70.2 +/- 5.4]%) than in the other abstinence groups (P < 0.01), so was it in the normal sperm morphology rate > 20% group ([71.4 +/- 5.1]%) than in the others (P < 0.01). The survival rate of the post-thaw sperm was not correlated with seasons, blood types, and semen volume (P > 0.05). CONCLUSION: Sperm motility, concentration and morphology and the abstinence period of sperm donors may be valuable predictors of the cryosurvival rate of post-thaw sperm, but no correlation was found between the survival rate and seasons, blood types and semen volume.

Efficacy of swim-up versus density gradient centrifugation in improving sperm deformity rate and DNA fragmentation index in semen samples from teratozoospermic patients.

PURPOSE: To compare the efficacy of swim-up and DGC in improving sperm deformity and DNA fragmentation and to determine which method is better in teratozoospermic patients requiring artificial reproduction. METHODS: The present study compared the effects of swim-up and density gradient centrifugation (DGC), the two most commonly used semen preparation methods, on sperm deformity rate and DNA fragmentation index (DFI) in semen samples from teratozoospermic patients. RESULTS: The results demonstrated that both swim-up and DGC yielded a significantly lower sperm deformity rate and DFI in comparison to unprocessed whole semen, with DGC having more favorable results. Sperm deformity rate in unprocessed whole semen samples was significantly lower in the 20-29 age group than in the 40-49 age group, but no significant difference was observed in DFI between different age groups. There was no significant correlation between sperm deformity rate and DFI. CONCLUSIONS: Our findings suggest that enrichment of sperm with normal morphology and intact DNA in teratozoospermic patients could be achieved by both DGC and swim-up procedures, and that DGC is a better method.

[Relationship between the expression of HBV mRNA in embryos and father-to-infant HBV transmission].

OBJECTIVE: To investigate father-to-infant transmission of hepatitis B virus (HBV) by detecting HBV mRNA in the IVF embryos with paternal HBV infection. METHODS: We collected 18 discarded IVF embryos (9 cases) with paternal chronic HBV infection, and detected HBV mRNA in the embryos by single-cell RT-PCR. RESULTS: HBV mRNA positive signals were found in 1 of the 18 embryos with paternal serum HBV positive markers (5.6%), but no specific HBV mRNA signals were observed in the 84 embryos of the negative control group. Follow-up visits revealed no significant difference between the experimental and negative control groups either in the rate of clinical pregnancy (P > 0.05) or in that of early abortion (P > 0.05). The IVF embryo with paternal HBV mRNA positive signals was successfully implanted, but early abortion occurred. HBV infection was not transmitted to progeny in either of the two groups. CONCLUSION: The positive results of HBV mRNA indicate that HBV can get into early-cleavage embryos through sperm and replicate there, which may be the main channel of father-to-infant transmission. HBV may interfere with the development of embryos, and even result in abortion and other adverse outcomes.

[Effect of domestic highly purified urinary follicle stimulating hormone on outcomes of in vitro fertilization-embryo transfer in controlled ovarian stimulation].

OBJECTIVE: To investigate the effect of domestic urine-derived high-purity follicle- stimulating hormone (HP-FSH, Lishenbao) on the outcome of in vitro fertilization(IVF) embryo transfer (ET) in controlled ovarian stimulation (COS). METHODS: From 1 September 2010 to 31 March 2011, total of 3178 infertility patients from 14 Reproductive Center with IVF or intracytoplasmic sperm injection (ICSI) indications who accepted first IVF or ICSI cycle were studied retrospectively. Their causes of infertility include all infertility factors except ovulatory dysfunction infertility and uterine factor infertility. The only long luteal phase gonadotropin-releasing hormone agonist (GnRH-a) protocol was included. Patients were divided into 2 groups according to the type of follicle-stimulating hormone (FSH) agents used: 1932 cases in HP-FSH group and 1246 cases in recombinant FSH (rFSH)group. Patients in both groups were combined with human menopausal gonadotropin (hMG) at doses of 150 U when follicle with diameter reached to 14-16 mm. When 3 dominate follicle with diameter reached 18 mm, hCG at dose of 5000 to 10 000 U or recombinant hCG at dose of 250 µg was administered by intramuscular injection. After 34 to 36 hours, oocytes were obtained guided by ultrasound, then IVF-ET were underwent in their Reproductive Center. The primary endpoint was comparison of live birth rate between the two groups. The secondary endpoints were comparisons of clinical pregnancy rate, miscarriage rate, and implantation rate, as well as COS and IVF outcome between the two groups. RESULTS: (1) There were significantly differences in baseline characteristics of the patients between two groups. The mean age was elder(32 ± 4 versus 30 ± 4, P < 0.01) , the infertility duration was longer (5 ± 4 versus 5 ± 3, P < 0.01) , and antral follicle count (AFC) was less (11 ± 5 versus 13 ± 7, P < 0.01) in patients of HP-FSH group compared with those in patients of rFSH group. (2) As compared with rFSH, the total doses of gonadotropin needed was (2348 ± 1011) U in HP-FSH group versus (2022 ± 659) U in rFSH group, the number of oocytes 13 ± 6 in HP-FSH group and 14 ± 7 in rFSH group, the rate of embryo frozen cycle of 66.30% (1281/1932) in HP-FSH group and 74.88% (933/1246) in rFSH group, which all reached statistical difference (P < 0.01). However, there were no significant different implantation rate [30.49% (1111/3644) versus 32.45% (737/2271)] between two groups. The other clinical parameters did not show significant difference, including clinical pregnancy rate per started cycle [41.61% (804/1932) versus 41.97% (523/1246) ] , clinical pregnancy rate per ET cycle[46.58% (804/1726) versus 48.47% (523/1079)], live birth rate per started cycle[34.21% (661/1932) versus 34.19% (426/1246)], live birth rate per ET cycle [38.30% (661/1726) versus 39.48% (426/1079)], miscarriage rate[13.6% (109/804) versus 16.4% (86/523)], and moderate/severe ovarian hyperstimulation syndrome (OHSS) rate [5.80% (112/1932) versus 7.78% (97/1246)](P > 0.05).(3) Treatment cost: the cost of gonadotropins needed for the patients in HP-FSH group was lower than that in rFSH group (4005 ± 1650 versus 6482 ± 2095, P < 0.01). CONCLUSION: In IVF/ICSI treatment cycles, domestic HP-FSH has similar live birth rate and lower financial burden when compared with rFSH.

[Expressions of sperm-specific genes in carnitine-cultured testis sperm of obstructive azoospermia patients].

OBJECTIVE: To investigate the effects of carnitine on human sperm motility and its potential role in the treatment of male infertility diseases. METHODS: We obtained sperm by testis puncture from obstructive azoospermia patients and cultured them in vitro with normal culture solution (the control group) and the solution with L-carnitine at the concentration of 100 and 250 mmol/L, respectively. We observed the changes in sperm motility and morphology before and after the treatment, detected the expressions of the germ-specific genes, Vasa, Dazl, Acr, Prm1 and ATPase 6.0 by RT-PCR, and investigated the relationship between L-carnitine and the genes associated with sperm development and maturation. RESULTS: After 24 -72 hours of treatment, the percentage of motile sperm was significantly higher in the 100 mmol/L L-carnitine group than in the control and 250 mmol/L L-carnitine groups (P < 0.01); the number of forward moving sperm was obviously increased and sperm morphology remained normal in the 100 mmol/L L-carnitine group. RT-PCR showed that L-carnitine increased the expressions of Acr, Prm1, Dazl and ATPase 6. 0 at the concentration of 100 mmol/L, and decreased the expressions of Dazl, Acr and Prm1 at 250 mmol/L. CONCLUSION: L-carnitine at a proper concentration may improve the motility of incubated testicular sperm by upregulating the expressions of some sperm-specific genes, which helps sperm selection for intracytoplasmic sperm injection. However, a higher concentration of L-carnitine may reduce the expressions of these genes, probably due to its increased toxicity.

[Teratozoospermia affects in vitro fertilization outcome].

OBJECTIVE: To analyze the possible causes of total fertilization failure (TFF) in conventional in vitro fertilization (IVF). METHODS: We included in this study 552 couples who accomplished the first conventional IVF cycle in our center from January 2007 to August 2008. All the males met the Kruger/Tygerberg criteria, with the teratozoospermia rate < 96% in the previous 12 months and the postwash motile sperm > 1 million on the day of egg retrieval. The eggs were fertilized totally by conventional IVF. RESULTS: Of the total number, 515 couples got zygotes by conventional IVF, with a fertilization rate of 78.67%, and 37 suffered TFF. The rates of primary infertility and teratozoospermia were significantly higher in the TFF group than in the fertilized group (P < 0.05). There were no statistically significant differences between the two groups in the number of controlled ovarian hyperstimulation (COH) days, the number of oocytes retrieved, the dose of gonadotropin (Gn) used, the age of the couples, the length of protocols and the rate of oligoasthenozoospermia. CONCLUSION: Intracytoplasmic sperm injection (ICSI) can be considered for at least some of the oocytes to avoid unnecessary fertilization failure in teratozoospermia patients by Kruger/Tygerberg strict criteria.